Murray H W, Spitalny G L, Nathan C F
J Immunol. 1985 Mar;134(3):1619-22.
To determine the role of IFN-gamma in the activation of resident mouse peritoneal macrophages, crude macrophage-activating lymphokines were incubated with a monoclonal anti-murine IFN-gamma antibody. This treatment abolished the capacity of mitogen-induced lymphokines to enhance either H2O2 release or activity against the intracellular protozoa Toxoplasma gondii and Leishmania donovani. All macrophage-activating factor detected by these assays was also removed by passing the lymphokines over a Sepharose column to which the monoclonal anti-IFN-gamma antibody had been coupled. Therefore, pure murine rIFN-gamma was tested both in vitro and in vivo as a single activating agent. After 48 hr of pretreatment in vitro with 0.01 to 1 antiviral U/ml, macrophage H2O2-releasing capacity was enhanced an average of 6.4-fold; half-maximal stimulation was induced by 0.03 U/ml. Resident macrophages infected with T. gondii half-maximally inhibited parasite replication after 24 hr of preincubation with 0.14 U/ml of rIFN-gamma, and near complete inhibition was achieved by pretreatment with 100 U/ml. Half-maximal leishmanicidal activity was induced by 0.08 U/ml of rIFN-gamma, and 67 to 75% of intracellular L. donovani amastigotes were killed after macrophages were preincubated with 10 to 100 U/ml. Eighteen hours after parenteral injection of rIFN-gamma, peritoneal macrophages displayed a dose-dependent enhancement of H2O2-releasing capacity and antiprotozoal activity. Half-maximal enhancement required 85 to 250 U or rIFN-gamma given i.p. Peritoneal macrophages were also activated by rIFN-gamma injected i.v. and intramuscularly. These results suggest that, in the mouse model, IFN-gamma is likely to be a primary factor within mitogen-induced lymphokines responsible for activating macrophage oxidative metabolism and antiprotozoal activity, and indicate that rIFN-gamma is a potent activator of these effector functions both in vitro and in vivo. These findings provide a rationale for evaluating rIFN-gamma in the treatment of systemic intracellular infections, and indicate that murine models are appropriate for such studies.
为确定γ干扰素在激活小鼠腹腔常驻巨噬细胞中的作用,将粗制巨噬细胞激活淋巴因子与单克隆抗小鼠γ干扰素抗体一起孵育。这种处理消除了丝裂原诱导的淋巴因子增强过氧化氢释放或针对细胞内原生动物刚地弓形虫和杜氏利什曼原虫活性的能力。通过使淋巴因子通过偶联有单克隆抗γ干扰素抗体的琼脂糖柱,这些检测中所检测到的所有巨噬细胞激活因子也被去除。因此,对纯的小鼠重组γ干扰素在体外和体内作为单一激活剂进行了测试。在体外以0.01至1抗病毒单位/毫升预处理48小时后,巨噬细胞过氧化氢释放能力平均提高了6.4倍;0.03单位/毫升诱导出半数最大刺激。在与0.14单位/毫升的重组γ干扰素预孵育24小时后,感染刚地弓形虫的常驻巨噬细胞对寄生虫复制的抑制达到半数最大,用100单位/毫升预处理可实现近乎完全抑制。0.08单位/毫升的重组γ干扰素诱导出半数最大杀利什曼原虫活性,在用10至100单位/毫升预孵育巨噬细胞后,67%至75%的细胞内杜氏利什曼原虫无鞭毛体被杀死。经腹腔注射重组γ干扰素18小时后,腹腔巨噬细胞的过氧化氢释放能力和抗寄生虫活性呈剂量依赖性增强。半数最大增强需要腹腔注射85至250单位的重组γ干扰素。静脉内和肌肉内注射重组γ干扰素也可激活腹腔巨噬细胞。这些结果表明,在小鼠模型中,γ干扰素很可能是丝裂原诱导的淋巴因子中负责激活巨噬细胞氧化代谢和抗寄生虫活性的主要因子,并表明重组γ干扰素在体外和体内都是这些效应功能的有效激活剂。这些发现为评估重组γ干扰素治疗全身性细胞内感染提供了理论依据,并表明小鼠模型适用于此类研究。